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Methods for high yield production of terpenes

Inactive Publication Date: 2017-01-03
DONALD DANFORTH PLANT SCI CENT
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

This approach allows for the first time the large-scale commercial production of terpenes in plants, providing a renewable source of aromatic hydrocarbons suitable for jet fuel, organic solvents, and industrial raw materials, with seed terpene content exceeding 1.0 mg / g dry weight.

Problems solved by technology

Camelina is not a foodstuff plant and grows on marginal lands (e.g. Montana) that are generally considered unsuitable for large scale food production.

Method used

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  • Methods for high yield production of terpenes
  • Methods for high yield production of terpenes
  • Methods for high yield production of terpenes

Examples

Experimental program
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example 1

Experimental Procedures

[0312]Plant Materials:

[0313]Wild-type Camelina sativa was grown in the green house at Donald Danforth Plant Science Center. Peppermint Mentha piperita leaves were harvested from a garden in St. Louis, Mo. in September, 2009.

[0314]Cloning of Peppermint Geranyl Diphosphate Synthase and Limonene Synthase cDNAs

[0315]Total RNA was extracted from peppermint leaves using RNeasy plant mini kit (Qiagen). First-strand cDNAs was synthesized using SuperScript III (Invitrogen). References and nucleotide sequences (Burke et al. (1999) Arch. Biochem. Biophys., 422, 52-60; Alonso et al. (1992) J Biol Chem., 267, 7582-7; Colby et al. (1993) J Biol Chem., 268, 23016-24. NCBI accession numbers: AF182827, AJ249453, EU108697, AW255818) were used to design cloning primers. Geranyl diphosphate synthase small subunit without predicted chloroplast transit peptide has been cloned from the peppermint cDNAs with primers: GSSfC and GSSr4 (Table E1) (FIG. 10). Geranyl diphosphate synthase ...

example 2

Biosynthesis and Accumulation of Cyclic Monoterpene Hydrocarbon (4S)-Limonene and the Bicyclic Sesquiterpene Hydrocarbon 5-Epi-Aristolochene in Camelina Seed

[0330]This example describes the biosynthesis and accumulation of the cyclic monoterpene hydrocarbon (4S)-limonene and the bicyclic sesquiterpene hydrocarbon 5-epi-aristolochene in camelina seed by expressing appropriate combinations of terpene biosynthetic enzymes.

[0331]The phrase “biosynthetically appropriate combination of enzymes” refers to a combination of terpene biosynthetic enzymes that facilitates the biosynthesis of a monoterpene or sesquiterpene of interest. Such combinations include a combination of: 1) a geranyl diphosphate synthase and a monoterpene synthase that catalyzes the formation of a monoterpene of interest, or 2) a combination of a farnesyl diphosphate synthase and a sesquiterpene synthase that catalyzes the formation of a sesquiterprene of interest. The phrase “a biosynthetically appropriate combination o...

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Abstract

Provided are enhanced high yield production systems for producing terpenes in plants via the expression of fusion proteins comprising various combinations of geranyl diphosphate synthase large and small subunits and limonene synthases. Also provided are engineered oilseed plants that accumulate monoterpene and sesquiterpene hydrocarbons in their seeds, as well as methods for producing such plants, providing a system for rapidly engineering oilseed crop production platforms for terpene-based biofuels.

Description

CROSS REFERENCE TO RELATED APPLICATIONS[0001]This application claims the benefit of priority of U.S. Provisional Application Ser. No. 61 / 645,877, filed May 11, 2012, the contents of which are herein incorporated by reference in their entirety.STATEMENT REGARDING FEDERALLY SPONSORED RESEARCH OR DEVELOPMENT[0002]This invention was made with government support under DOE grant # DE-SC0001295 awarded by the Department of Energy. The government has certain rights in the invention.BACKGROUND OF THE INVENTION[0003]1. Field of the Invention[0004]The present invention relates to the enhanced production and accumulation of terpenes in plants via the expression of fusion proteins comprising various combinations of geranyl diphosphate synthase large and small subunits with limonene synthase. The present invention also relates to engineering of oilseed plants, exemplified by camelina, to accumulate monoterpene and sesquiterpene hydrocarbons, exemplified herein by the cyclic monoterpene hydrocarbo...

Claims

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Application Information

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Patent Type & Authority Patents(United States)
IPC IPC(8): A01H5/10C12N15/82C12N9/10C07C13/47C07C13/20C07C15/02C12N9/88C07C11/02C07C11/21
CPCC12N15/8257A01H5/10C07C11/02C07C11/21C07C13/20C07C13/47C07C15/02C12N9/1085C12N9/88C12N15/8243C07C2101/16C07K2319/08C07C2601/16
Inventor KUTCHAN, TONIHIGASHI, YASUHIROFENG, XIAOHONG
Owner DONALD DANFORTH PLANT SCI CENT
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